Hybrid adenovirus/adeno-associated virus vectors and methods of use thereof

Abstract

The invention provides recombinant vectors including adenovirus/adeno-associated virus (Ad/AAV) vectors and mini-adenovirus (mAd) vectors. Further, the invention provides cells containing these vectors, and methods for making and using the vectors and cells. The compositions and methods of the invention are useful in transferring nucleotide sequences of interest into a cell, including, but not limited to, in gene therapy applications.

1. A recombinant vector, comprising in operable combination:
 a) an adeno-associated virus terminal repeat-DD (AAV TR-DD) sequence; 
 b) first and second inverted copies of a nucleotide sequence of interest having a 5′ end and a 3′ end and flanking said AAV TR-DD sequence; 
 c) left and right inverted terminal repeats (ITRs) of an adenovirus flanking said first and second inverted copies of said nucleotide sequence of interest; 
 d) a first adenovirus packaging sequence operably linked to one of said ITRs; and 
 e) one or more adenovirus genes. 
 
     
     
       2. The vector of  claim 1 , wherein said packaging sequence is linked to said 5′ end or said 3′ end of said nucleotide sequence of interest. 
     
     
       3. The vector of  claim 1 , wherein said nucleotide sequence of interest comprises an adeno-associated virus rep gene region. 
     
     
       4. A recombinant vector, comprising in operable combination:
 a) an adeno-associated virus terminal repeat-DD (AAV TR-DD) sequence; 
 b) first and second inverted copies of a nucleotide sequence of interest having a 5′ end and a 3′ end and flanking said AAV TR-DD sequence; 
 c) left and right inverted terminal repeats (ITRs) of an adenovirus flanking said first and second inverted copies of said nucleotide sequence of interest; and 
 d) a first adenovirus packaging sequence operably linked to one of said ITRs, wherein said vector is a gutted adenovirus vector. 
 
     
     
       5. The vector of  claim 1 , wherein said one or more adenovirus genes is selected from an E1, an E2, an E3, and an E4 gene region. 
     
     
       6. The vector of  claim 5 , wherein said vector lacks an E1 gene region. 
     
     
       7. The vector of  claim 6 , wherein said vector further lacks an E3 gene region. 
     
     
       8. The vector of  claim 5 , wherein said vector lacks an E3 gene region. 
     
     
       9. The vector of  claim 5 , wherein said vector lacks an E4 gene region. 
     
     
       10. The vector of  claim 5 , wherein said vector lacks an E2 gene region. 
     
     
       11. The vector of  claim 1 , wherein said vector further comprises e) first and second inverted adeno-associated virus terminal repeat D sequences flanking said first and second inverted copies of said nucleotide sequence of interest, wherein said first and second inverted adeno-associated virus terminal repeat D sequences are flanked by said left and right inverted terminal repeats (ITRs) of adenovirus. 
     
     
       12. The vector of  claim 1 , wherein said vector further comprises a second adenovirus packaging sequence linked to one of said inverted terminal repeats (ITRs). 
     
     
       13. The vector of  claim 1 , wherein said vector further comprises (e) first and second inverted adeno-associated virus terminal repeat D sequences flanking said first and second inverted copies of said nucleotide sequence of interest, wherein said first and second inverted adeno-associated virus terminal repeat D sequences are flanked by said left and right inverted terminal repeats (ITRs) of adenovirus, and (f) a second adenovirus packaging sequence linked to one of said inverted terminal repeats (ITRs). 
     
     
       14. A cell comprising the recombinant vector of  claim 1 , wherein said cell is selected from the group consisting of a (a) cell in vitro, and (b) cell in vivo in a non-human animal. 
     
     
       15. The cell of  claim 14 , wherein said cell lacks expression of one or more adenovirus early gene regions selected from the group consisting of an E1, an E2, an E3, and an E4 gene regions. 
     
     
       16. The cell of  claim 14 , wherein said cell comprises a primary cell. 
     
     
       17. The cell of  claim 16 , wherein said primary cell is selected from the group consisting of a mouse cell and a human cell. 
     
     
       18. The cell of  claim 14 , wherein said cell comprises a cell line. 
     
     
       19. The cell of  claim 18 , wherein said cell line is selected from the group consisting of a HeLa cell line, a A549-derived cell line, a 293-derived cell line, a HepG2-derived cell line, a COS1-derived cell line, a HMEC-derived cell line, a KB-derived cell line, a JW-22-derived cell line, a Neo6-derived cell line and a C12-derived cell line.